林文澧臺灣大學:醫學工程學研究所呂國維Lu, Kuo-WeiKuo-WeiLu2010-05-182018-06-292010-05-182018-06-292009U0001-2707200917032600http://ntur.lib.ntu.edu.tw//handle/246246/183723巨分子藥物可以標定腫瘤進行專一性治療，且有較長的血液半衰期，是現今腫瘤藥物主要發展的對象，但因為腫瘤組織本身特性的限制，使得巨分子藥物無法進入腫瘤組織內部，要克服此問題，除了從藥物的改良著手外，改變腫瘤組織的特性也是一個方法。前人研究指出聚焦式超音波搭配超音波顯影劑可使微血管壁破裂，特定參數的使用已經證明可讓藥物在活體中遞送達到更好的效果。研究主要探討聚焦式超音波搭配顯影劑造成血管通透性的改變。我們使用螢光代糖當作巨分子藥物載體，並以雙光子顯微技術活體且動態觀察螢光代糖在小鼠正常血管的滲透情形。超音波的頻率為1MHz，最大負聲壓0.6MPa，重覆頻率為1Hz，脈衝長度10ms。 實驗結果得知，超音波搭配顯影劑可破壞正常組織的血管壁，並造成巨分子從血管中滲出。並藉由Nakamura等人在1974年所建立的分子運輸模型與本研究來做數據分析，計算出施打超音波之後巨分子在組織的運輸參數，並進一步由文獻探討得知，超音波搭配超音波顯影劑會造成巨分子在血管外的擴散係數增加。此種研究方法可用於研究超音波搭配顯影劑對於腫瘤組織的影響，並提供腫瘤治療的策略。Macromolecular drug carrier is one of the major research subjects in anti-cancer drug development today due to its long half-life and specific targeting ability. However, its penetration ability into the tumor tissue is constrained by the tumor’s microenvironments. To improve the anti-cancer drug distribution in the tumor tissue, we can modify the drug’s functions and characteristics or change the tumor’s vascular properties and microenvironments. It has been shown that focused ultrasound with microbubbles could disrupt the vascular wall and the use of specific parameters could enhance the drug delivery. In this study, we investigated the permeability variation of blood vessels into tissue for focused ultrasound sonication in the presence of microbubbles (ultrasound contrast agent). We used dextran rhodamine as micromolecular drug carrier and used two-photon microscope to observe the permeation of dextran from the blood vessels into the tissue of normal mouse. The focused ultrasound used is 1.0 MHz driving frequency, 0.6 MPa peak negative pressure, 1 Hz repetition frequency, and 10 ms burst, and the ultrasound contrast agent dose was 200 μL/kg injected into the mouse tail vein. The experimental results showed that focused ultrasound with microbubbles can effectively disrupt the blood vessel walls of normal mouse tissue and cause the extravasation of macromolecules. We used a mathematical model with diffusion (Nakamura et al.,1974) to calculate and analyze the transport parameters of macromolecules during ultrasound sonication. We found that the diffusion coefficient of macromolecules during sonication was greater than that published in literatures. The methods used in this study can further be employed to investigate the transport characteristics of tumor blood vessels and the dose distribution in tumor tissue for focused ultrasound with microbubbles, and the results may provide useful information for future cancer treatment strategy.誌謝..............................................................I要.............................................................IIbstract........................................................III錄.............................................................IV目錄...........................................................VI目錄..........................................................VII1章 緒論......................................................1.1 化學治療..................................................1.2 藥物遞送(Drug Delivery)...................................1.2.1 藥物運輸(Transport)與遞送原理.............................1.2.2 生物組織特性與藥物遞送....................................2.3 巨分子藥物載體(Macromolecular Drug Carrier)...............3.4 超音波與生物物質的交互作用................................4.4.1 機械效應..................................................4.4.2 熱效應....................................................4.4.3 空蝕化效應(Cavitation)....................................5.5 超音波促進藥物遞送........................................5.6 研究目的..................................................62章 實驗材料、設備與方法......................................9.1 實驗材料與設備................................................9.1.1 實驗動物..................................................9.1.2 皮膚固定裝置..............................................9.1.3 螢光代糖(Dextran).........................................9.1.4 雙光子顯微系統...........................................10.1.5 超音波設備...............................................11.2 實驗方法.....................................................12.2.1 超音波設備與雙光子系統的結合.............................12.2.2 超音波參數...............................................12.2.3 實驗流程.................................................13.2.4 血管外影像分析...........................................13.3 理論分析.....................................................143章 結果.....................................................26.1 柱狀型聚焦式超音波探頭特性...................................26.2 小鼠正常血管時間間隔(Time-Lapse)影像.........................26.3 影像中螢光訊號初步分析.......................................27.3.1 未施打超音波血管外螢光強度隨時間的變化...................27.3.2 施打超音波時血管外螢光強度隨時間的變化...................27.3.3 施打超音波後血管外螢光強度隨時間的變化...................27.4 螢光訊號結合理論分析求得分子運輸參數.........................274章 討論.....................................................40.1 螢光強度偵測.................................................40.2 超音波搭配顯影劑使螢光代糖滲出到微血管外.....................40.3 分子擴散係數.................................................41.3.1 超音波搭配顯影劑對於分子擴散係數的影響...................41.3.2 數據取得、處理及計算.....................................42.3.3 數學模型.................................................435章 結論與未來展望...........................................46考文獻.........................................................47application/pdf1141667 bytesapplication/pdfen-US聚焦式超音波超音波顯影劑雙光子顯微術血管運輸參數Focused ultrasoundUltrasound contrast agentTwo-photon microscopeBlood vesselsTransport parameters[SDGs]SDG3thesishttp://ntur.lib.ntu.edu.tw/bitstream/246246/183723/1/ntu-98-R96548046-1.pdf